Neutral Lipid Classes Research Articles

Formation and mobilization of neutral lipids in the yeast Saccharomyces cerevisiae

Since energy storage is a basic metabolic process, the synthesis of neutral lipids occurs in all kingdoms of life. The yeast Saccharomyces cerevisiae, widely accepted as a model eukaryotic cell, contains two classes of neutral lipids, namely STEs (steryl esters) and TAGs (triacylglycerols). TAGs are synthesized through two pathways governed by the acyl-CoA diacylglycerol acyltransferase Dga1p and the phospholipid diacylglycerol acyltransferase Lro1p. STEs are formed by two STE synthases Are1p and Are2p, two enzymes with overlapping function, which also catalyse TAG formation, although to a minor extent. Neutral lipids are stored in the so-called lipid particles and can be utilized for membrane formation under conditions of lipid depletion. For this purpose, storage lipids have to be mobilized by TAG lipases and STE hydrolases. A TAG lipase named Tgl3p was identified as a major yeast TAG hydrolytic enzyme in lipid particles. Recently, a new family of hydrolases was detected which is required for STE mobilization in S. cerevisiae. These enzymes, named Yeh1p, Yeh2p and Tgl1p, are paralogues of the mammalian acid lipase family. The role of these proteins in biosynthesis and mobilization of TAG and STE, and the regulation of these processes will be discussed in this minireview.

Qualitative and Quantitative Analysis of Lipid Classes in Fish Oils by Thin‐Layer Chromatography with an Iatroscan Flame Ionization Detector (TLC‐FID) and Liquid Chromatography with an Evaporative Light Scattering Detector (LC‐ELSD)

Combined effects of hydrogen and air flow rates on the peak response of selected neutral lipid classes (triacylglycerol, diacylglycerol, monoacylglycerol, free fatty acids, and ethyl esters) were studied to optimize and calibrate the Iatroscan Mk‐6s Chromarod system for the qualitative and quantitative analysis of lipid classes by thin‐layer chromatography (TLC) with flame ionization detection in fish oil during the transesterification process. Air flow rate of 2 L/min, hydrogen flow rate of 150–160 mL/min, and scan rate of 30 s/rod were found to be the optimum conditions. All samples were also analyzed by high performance liquid chromatography (HPLC) with evaporative light scattering detection. Quantitative results obtained by TLC with the flame ionization detection method were comparable to those obtained from HPLC with evaporative light scattering detection.

Characterisation of fatty acids and bioactive compounds of kachnar ( Bauhinia purpurea L.) seed oil

Information concerning the exact composition of kachnar ( Bauhinia purpurea) seed oil is scare. In the present contribution, a combination of CC, GC, TLC and normal-phase HPLC were performed to analyse lipid classes, fatty acids and fat-soluble bioactives of kachnar seed oil. n-Hexane extract of kachnar oilseeds was found to be 17.5%. The amount of neutral lipids in the crude seed oil was the highest (ca. 99% of total lipids), followed by glycolipids and phospholipids, respectively. Linoleic, followed by palmitic, oleic and stearic, were the major fatty acids in the crude seed oil and its lipid classes. The ratio of unsaturated fatty acids to saturated fatty acid, was higher in neutral lipid classes than in the polar lipid fractions. The oil was characterised by a relatively high amount of phytosterols, wherein the sterol markers were β-sitosterol and stigmasterol. β-Tocopherol was the major tocopherol isomer with the rest being δ-tocopherol. In consideration of potential utilisation, detailed knowledge of the composition of kachnar ( B. purpurea) seed oil is of major importance.

기능성 지질 추출 소재로서 Fish Frames의 지질성분 특성

수산가공 부산물인 fish frame을 기능성 지질 추출소재로서의 이용 가능성을 검토하기 위하여 6종의 fish frame(붉은 메기, 연어, 삼치, 고등어, 붕장어, 가다랑어 등)에 대한 지질 성분 특성에 대하여 살펴보았다. Fish frame에 대한 어류 뼈의 수율은 가다랑어가 64.2%로 가장 높았고, 다음으로 연어(57.9%), 붕장어(54.6%), 붉은 메기(42.0%), 삼치(41.7%)등의 순이었으며, 고등어가 32.6%로 가장 낮았다. 어류 뼈 유래 총지질 중 중성지질의 함량은 삼치가 23.3 g/100 g으로 가장 많았고, 다음으로 붕장어(21.5 g/100 g), 연어(16.0 g/100 g), 고등어(15.5 g/100 g)의 순이었으며, 가다랑어와 붉은 메기의 경우 다른 4종의 어류에 비하여 확연히 낮았다. 붉은 메기를 제외한 5종 어류의 중성지질 모두가 근육 및 뼈 지방에 관계없이 TG가 주성분(근육지방, 71.9~83.2%; 뼈지방, 74.2~86.9%)이었고, 다음으로 근육지방의 경우, FFA(8.1~19.2%), 뼈지방의 경우 FS(5.5~15.5%)이었다. 붉은 메기의 경우, 중성지질의 주성분인 TG의 조성비(근육지방, 48.6%; 뼈지방, 45.3%)가 다른 어류에 비하여 확연히 낮았고, FFA의 조성비 또한 근육지방(41.6%) 및 뼈지방 (43.4%)에 관계없이 모두 5종의 어류에 비하여 확연히 높았다. 어류 뼈의 중성지질의 EPA＋DHA 조성은 연어가 29.3%로 가장 높았고, 다음으로 가다랑어(27.1%), 붉은 메기(27.0%), 고등어(25.7%), 삼치(21.6%)의 순이었고, 붕장어의 경우 14.9%에 불과하였다. 이상의 수율, 중성지질 함량, 유리지방산 조성 및 EPA＋DHA 조성 비율의 결과로 미루어 보아 기능성 지질 추출원으로는 연어가 가장 우수하였고, 다음으로 삼치, 붕장어, 고등어, 가다랑어 등의 순이었으며, 붉은 메기의 경우 부적절하다고 판단되었다.

Lipid analysis of the plasma membrane and mitochondria of brewer’s yeast

The plasma membrane and mitochondria of bottom fermenting brewer's yeast obtained as a by-product of industrial beer production were isolated and the lipid fraction was analyzed. The phospholipid content accounted for 78 mg/g protein in the plasma membrane and 59 mg/g protein in the mitochondria. Major phospholipids in both preparations were phosphatidylinositol, phosphatidylcholine and phosphatidylethanolamine but their proportions differed significantly. In the plasma membrane phosphatidylinositol, and in the mitochondria phosphatidylcholine were present in the highest concentration (37 and 30%, respectively). The main classes of neutral lipids (triacylglycerols, ergosterol, squalene and steryl esters) were twice more abundant in the plasma membrane than in the mitochondria (61 and 33 mg/g protein, respectively). A characteristic of the neutral lipid composition of both organelles was the low content of ergosterol (12 and 7 mg/g protein, respectively) and a high content of squalene (25 and 22 mg/g protein). The main feature of the fatty acid composition of both organelles was the preponderance of saturated fatty acids (78 and 79%, respectively), among which palmitic acid was the principal one. The most expressed characteristics of lipid fractions of the analyzed plasma membranes and mitochondria, high concentration of squalene and preponderance of saturated fatty acids are the consequences of anaerobic growth conditions. The lack of oxygen had possibly the strongest effect on the lipid composition of the plasma membranes and mitochondria of bottom fermenting brewer's yeast.

Changes in amino acids and lipids during embryogenesis of European lobster, Homarus gammarus (Crustacea: Decapoda)

We studied the amino acid and lipid dynamics during embryogenesis of Homarus gammarus. Major essential amino acids (EAA) in the last stage of embryonic development were arginine, lysine and leucine; major nonessential amino acids (NEAA) were glutamic acid, aspartic acid, valine and glycine. The highest percent of utilization occurred in respect to EAA (27.8%), mainly due to a significant decrease ( p<0.05) of methionine (38.3%) and threonine (36.0%). NEAA also decreased significantly ( p<0.05, 11.4%), namely serine (38.1%), tyrosine (26.4%) and glutamic acid (25.7%). In contrast, the free amino acid content increased significantly ( p<0.05) during embryonic development, especially the free nonessential amino acids (FNEAA). In the last stage, the most abundant FNEAA were glycine, proline, alanine and taurine, and the major free essential amino acids (FEAA) were arginine, lysine and leucine. Lipid content decreased significantly ( p<0.05) during embryonic development. A substantial decrease in all neutral lipid classes was observed (>80% of utilization). Major fatty acids were 16:0, 18:0, 18:1n-9, 18:2n-6, 18:3n-3, 20:5n-3 and 22:6n-3. Unsaturated (UFA) and saturated fatty acids (SFA) were used up at similar rates (76.5% and 76.3%, respectively). Within UFA, monounsaturates (MUFA) were consumed more than polyunsaturates (PUFA) (82.9% and 67.5%, respectively).

Composition and seasonal variation of fatty acids of Diplodus vulgaris L. from the Adriatic Sea

AbstractMuscle tissue from the common two‐banded sea bream Diplodus vulgaris L. originating from the Adriatic Sea, Croatia, was analyzed. The FA composition of neutral (TAG) and polar (PE, PC, PI/PS) lipid classes was determined, as well as the lipid and water contents during winter and summer periods. Both the total lipid and water contents were higher in the winter period. We identified 16 different FA. The major constituents of the total FA in both seasons were saturates: palmitic (16∶0) and stearic acids (18∶0); monoenes: oleic (18∶1n−9) and palmitoleic acids (16∶1n−7); and polyunsaturates: arachidonic acid (20∶4n−6), EPA (20∶5n−3), and DHA (22∶6n−3), but their amounts and ratios differed significantly between the two seasons and between lipid fractions. The FA composition showed a noticeable pattern of seasonality that reflected fluctuations mainly in TAG. The diminution of the monounsaturated FA content in the summer was clearly followed by an increase in PUFA content. Diplodus vulgaris is a good source of natural n−3 PUFA and would therefore be suitable for inclusion in highly unsaturated low‐fat diets.

Dynamics of neutral lipid storage in yeast.

Since energy storage is a basic metabolic process, the synthesis of neutral lipids occurs in all kingdoms of life. The yeast, Saccharomyces cerevisiae, widely accepted as a model eukaryotic cell, contains two classes of neutral lipids, namely steryl esters and triacylglycerols. Triacylglycerols are synthesized through two pathways governed by the acyl-CoA diacylglycerol acyltransferase Dga1p and the phospholipid diacylglycerol acyltransferase Lro1p, respectively. Steryl esters are formed by the two steryl ester synthases Are1p and Are2p, two enzymes with overlapping function which also catalyze triacylglycerol formation, although to a minor extent. Storage of neutral lipids is tightly linked to the biogenesis of so called lipid particles. The role of this compartment in lipid homeostasis and its interplay with other organelles involved in neutral lipid dynamics, especially the endoplasmic reticulum and the plasma membrane, are subject of current investigations. In contrast to neutral lipid formation, mobilization of triacylglycerols and steryl esters in yeast are less characterized at the molecular level. Only recently, the triacylglycerol lipase Tgl3p was identified as the first yeast enzyme of this kind by function. Genes and gene products governing steryl ester mobilization still await identification. Besides biochemical properties of enzymes involved in yeast neutral lipid synthesis and degradation, regulatory aspects of these pathways and cell biological consequences of neutral lipid depletion will be discussed in this minireview.

Dynamics of neutral lipid storage in yeast.

Since energy storage is a basic metabolic process, the synthesis of neutral lipids occurs in all kingdoms of life. The yeast, Saccharomyces cerevisiae, widely accepted as a model eukaryotic cell, contains two classes of neutral lipids, namely steryl esters and triacylglycerols. Triacylglycerols are synthesized through two pathways governed by the acyl-CoA diacylglycerol acyltransferase Dga1p and the phospholipid diacylglycerol acyltransferase Lro1p, respectively. Steryl esters are formed by the two steryl ester synthases Are1p and Are2p, two enzymes with overlapping function which also catalyze triacylglycerol formation, although to a minor extent. Storage of neutral lipids is tightly linked to the biogenesis of so called lipid particles. The role of this compartment in lipid homeostasis and its interplay with other organelles involved in neutral lipid dynamics, especially the endoplasmic reticulum and the plasma membrane, are subject of current investigations. In contrast to neutral lipid formation, mobilization of triacylglycerols and steryl esters in yeast are less characterized at the molecular level. Only recently, the triacylglycerol lipase Tgl3p was identified as the first yeast enzyme of this kind by function. Genes and gene products governing steryl ester mobilization still await identification. Besides biochemical properties of enzymes involved in yeast neutral lipid synthesis and degradation, regulatory aspects of these pathways and cell biological consequences of neutral lipid depletion will be discussed in this minireview.

Determination of c9,t11-CLA in major human plasma lipid classes using a combination of methylating methodologies.

Isomeric CLA exhibit several significant biological activities in animals and humans and are easily isomerized to their corresponding t,t-CLA isomers during methylation with various acid-catalyzed reagents. To minimize such isomerization and provide a valid quantification of human plasma CLA content, several methylation methods were tested. Plasma neutral lipid, nonesterified FA (NEFA), and polar lipid classes were separated into the following fractions: (i) cholesteryl ester (CE, 1.2 mg/12 mL, 37.5% lipids), (ii) TAG (0.8 mg/12 mL, 25% lipids), (iii) NEFA (0.2 mg/12 mL, 6.2% lipids), (iv) MAG/DAG/cholesterol (0.3 mg/12 mL, 9.4% lipids), and (v) phospholipid (PL, 0.5 mg/20 mL, 15.6% lipids). Data showed that c9,t11-CLA found in TAG, MAG/DAG/cholesterol, and PL fractions were converted to methyl esters with sodium methoxide within 2 h at 55 degrees C. However, the c9,t11-CLA in the CE fraction could not be completely converted to methyl esters by sodium methoxide/acetylchloride in methanol or methanolic KOH; instead, CE was treated with sodium methoxide and methyl acetate in diethyl ether for 1 h. NEFA were converted to methyl esters with trimethylsilyldiazomethane (TMSDAM). All reaction mixtures were monitored by TLC prior to GLC analysis. The highest enrichment of c9,t11-18:2 (% FA) was in TAG (0.31%), followed by CE (0.14%) and PL (0.13%). The above methylation methods were then applied to a small subset (n = 10) of nonfasting plasma lipid fractions to confirm the applicability of these data. Results from this subset of samples also indicated that the greatest enrichment of c9,t11-CLA was present in the TAG fraction (0.39%), followed by CE (0.27%) and PL (0.22%). These data indicate that different plasma fractions have different c9,t11-CLA contents.

Biochemical changes during the embryonic development of Norway lobster, Nephrops norvegicus

The objectives of the present study were to investigate the total and free amino acid profiles and lipid dynamics (lipid classes and fatty acids) during embryogenesis of Nephrops norvegicus , in order to understand the early larval protein and lipid requirements. There was a significant increase in total essential (EAA) and nonessential amino acid (NEAA) contents during embryonic development ( P <0.05). The major EAA were arginine, histidine and leucine, while the most important NEAA were glutamic acid, aspartic acid, glycine and proline. The higher percent increase occurred in respect to NEAA (19.9%), mainly due to the significant increase of glycine (51.8%) and alanine (35.1%). The free amino acid (FAA) content, especially the free nonessential amino acids (FNEAA), also increased significantly, being the quantitatively most important glycine, proline, taurine and glutamic acid. The free essential amino acids (FEAA) also revealed significant variations and the most important were arginine, lysine and leucine. A higher percent increase was attained with FNEAA (66.8%) in comparison to FEAA (49.6%), mainly due to the significant increase of homocystine (87.7%), valine (83.5%), glutamine (82.5%) and glutamic acid (76.1%). The absorption of dissolved organic compounds from seawater can explain the increase in free and total amino acid (FAA and TAA) contents, because a significant increase in the water content during development was observed ( P <0.05). A substantial decrease in all neutral lipid classes ( P <0.05) was observed during embryonic development, namely diacylglycerols (DAG) (97.9% of utilization), triacylglycerols (TAG) (93.2%), sterol esters (StE) (91.3%) and monoacylglycerols (MAG) (90.4%). The quantitatively most important fatty acids were the saturates (SFA) 14:0, 16:0 and 18:0, the monounsaturates (MUFA) 16:1n-7, 18:1n-9 and 18:1n-7, and the polyunsaturates (PUFA) 18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3. The unsaturated fatty acids (UFA) are used up at a higher rate (54.5% of utilization) than SFA (42.8%); within the UFA, MUFA are more consumed than PUFA (59.3% and 52.7%, respectively). In terms of the utilization of individual fatty acids, there was a preferential consumption of 20:4n-3, 22:6n-3, 18:1n-9, 22:5n-3 and 16:1n-7. It is evident that N. norvegicus depends primarily on lipid reserves of the egg during early ontogeny. On the contrary, this species tends to conserve EAA and increase NEAA and FNEAA contents during embryonic development.

4P-0942 Effects of medical nutrition therapy on the blood lipid levels in hyperlipidemic patients according to genetic polymorphism of Apo E

We studied the amino acid and lipid dynamics during embryogenesis of Homarus gammarus. Major essential amino acids (EAA) in the last stage of embryonic development were arginine, lysine and leucine; major nonessential amino acids (NEAA) were glutamic acid, aspartic acid, valine and glycine. The highest percent of utilization occurred in respect to EAA (27.8%), mainly due to a significant decrease (p<0.05) of methionine (38.3%) and threonine (36.0%). NEAA also decreased significantly (p<0.05, 11.4%), namely serine (38.1%), tyrosine (26.4%) and glutamic acid (25.7%). In contrast, the free amino acid content increased significantly (p<0.05) during embryonic development, especially the free nonessential amino acids (FNEAA). In the last stage, the most abundant FNEAA were glycine, proline, alanine and taurine, and the major free essential amino acids (FEAA) were arginine, lysine and leucine. Lipid content decreased significantly (p<0.05) during embryonic development. A substantial decrease in all neutral lipid classes was observed (>80% of utilization). Major fatty acids were 16:0, 18:0, 18:1n-9, 18:2n-6, 18:3n-3, 20:5n-3 and 22:6n-3. Unsaturated (UFA) and saturated fatty acids (SFA) were used up at similar rates (76.5% and 76.3%, respectively). Within UFA, monounsaturates (MUFA) were consumed more than polyunsaturates (PUFA) (82.9% and 67.5%, respectively).

4P-0944 Lipid levels are similar in patients on mediterranean and low fat diets for up to 3 years in patients with coronary heart disease on station therapy

The objectives of the present study were to investigate the total and free amino acid profiles and lipid dynamics (lipid classes and fatty acids) during embryogenesis of Nephrops norvegicus, in order to understand the early larval protein and lipid requirements. There was a significant increase in total essential (EAA) and nonessential amino acid (NEAA) contents during embryonic development (P<0.05). The major EAA were arginine, histidine and leucine, while the most important NEAA were glutamic acid, aspartic acid, glycine and proline. The higher percent increase occurred in respect to NEAA (19.9%), mainly due to the significant increase of glycine (51.8%) and alanine (35.1%). The free amino acid (FAA) content, especially the free nonessential amino acids (FNEAA), also increased significantly, being the quantitatively most important glycine, proline, taurine and glutamic acid. The free essential amino acids (FEAA) also revealed significant variations and the most important were arginine, lysine and leucine. A higher percent increase was attained with FNEAA (66.8%) in comparison to FEAA (49.6%), mainly due to the significant increase of homocystine (87.7%), valine (83.5%), glutamine (82.5%) and glutamic acid (76.1%). The absorption of dissolved organic compounds from seawater can explain the increase in free and total amino acid (FAA and TAA) contents, because a significant increase in the water content during development was observed (P<0.05). A substantial decrease in all neutral lipid classes (P<0.05) was observed during embryonic development, namely diacylglycerols (DAG) (97.9% of utilization), triacylglycerols (TAG) (93.2%), sterol esters (StE) (91.3%) and monoacylglycerols (MAG) (90.4%). The quantitatively most important fatty acids were the saturates (SFA) 14:0, 16:0 and 18:0, the monounsaturates (MUFA) 16:1n-7, 18:1n-9 and 18:1n-7, and the polyunsaturates (PUFA) 18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3. The unsaturated fatty acids (UFA) are used up at a higher rate (54.5% of utilization) than SFA (42.8%); within the UFA, MUFA are more consumed than PUFA (59.3% and 52.7%, respectively). In terms of the utilization of individual fatty acids, there was a preferential consumption of 20:4n-3, 22:6n-3, 18:1n-9, 22:5n-3 and 16:1n-7. It is evident that N. norvegicus depends primarily on lipid reserves of the egg during early ontogeny. On the contrary, this species tends to conserve EAA and increase NEAA and FNEAA contents during embryonic development.

Content and composition of phosphoglycerols and neutral lipids at different developmental stages of the eggs of the codling moth, Cydia pomonella (Lepidoptera: Tortricidae).

Phosphoglycerol, triacylglycerol, diacylglycerol, and free fatty acid content was studied in eggs of the codling moth Cydia pomonella at the white, red ring, and black head developmental stages. The composition of total phosphoglycerols and of the three classes of neutral lipids was also analyzed. The highest total lipid content was found in eggs at the white stage, the amount decreasing during development mainly as a result of a diminution in the quantity of phosphoglycerols, which account for approximately 50% of total content at all stages of egg development. The amount of triacylglycerols and free fatty acids changes significantly during development, whereas only minor changes were found in diacyglycerol levels. The total phosphoglycerol acyl composition of eggs at the white and red ring stages is similar, whereas differences are evident at the black head stage of development. Triacylglycerols and free fatty acids are enriched in saturated fatty acids in all analyzed stages. The acyl profile of diacylglycerols is different at each stage. The unsaturation index decreases in diacylglycerols and free fatty acids as a function of egg development. The results of the present paper suggest that triacylglycerols may constitute an important source of energy during the final period of egg development while phosphoglycerols may function as fuel during the beginning. Phosphoglycerols could be precursors for the triacylglycerol biosynthesis that takes place between white and red ring stages.

Lipid metabolism in the moss Rhytidiadelphus squarrosus (Hedw.) Warnst. from lead-contaminated and non-contaminated populations.

Lipid metabolism and the effect of Pb(2+) and Cu(2+) on this process was studied in the moss Rhytidiadelphus squarrosus collected from both a lead-contaminated and a non-contaminated site. Total radiolabelling of lipids from [1-(14)C]acetate was similar in both populations and Cu or Pb (1 microM, 10 microM) did not cause much alteration in acute exposure experiments. However, there were significant qualitative changes. Of the major labelled neutral lipid classes, samples from the lead-polluted site showed a decrease in labelling of triacylglycerols and an increase in wax esters. Acute lead exposure caused similar effects. Cu caused a decrease in the labelling of wax esters and an increase in diacylglycerols. These data suggest that heavy metals cause a change in carbon flux through the acylation reactions associated with the Kennedy pathway. R. squarrosus obtained from the Pb-contaminated site also showed changes in polar lipid labelling compared to the uncontaminated site. The labelling of phosphatidylcholine was more than halved and replaced by increased labelling of other zwitterionic lipids. The chloroplast glycerolipids were also increasingly labelled. Acute exposure to Pb, however, caused little alteration of labelling patterns within 24 h. R. squarrosus contains high levels of polyunsaturated fatty acids (PUFAs), but moss obtained from the Pb-polluted site had significantly less PUFAs containing three or more double bonds. Such samples, when incubated with [1-(14)C]acetate also showed decreased PUFA labelling. By contrast, acute exposure to Pb produced different results. These data provide a foundation for examining lipid metabolism in bryophytes and the effects of pollution in this important class of organism. The results also emphasize that acute and chronic exposure to heavy metals may produce different effects and that caution must be exercised in extrapolating data from one system to another.

Neutral lipid classes of black cumin (Nigella sativa L.) seed oils

n-Hexane (H) and chloroform:methanol (CM) were used to extract the oils of Nigella sativa L. seeds. A combination of column and thin-layer chromatography procedures on silica gel were performed to fractionate the main neutral lipid classes of seed oils. The fatty acid pattern of neutral lipid (NL) fractions, triacylglycerol (TAG) and sterol content were determined. NLs constituted 97.2% and 96.1% of total H and CM extract, respectively. TAGs were the major neutral lipid class (80.8–83.1% of the total NLs), while the NL profile was characterized by high levels of free fatty acids (14.3–16.2% of the total NLs). Linoleic acid (C18:2) was the predominant fatty acid followed by oleic (C18:1) and palmitic (C16:0) acids in all examined classes. A good resolution of the TAG critical pairs was accomplished employing a gas liquid chromatograph equipped with a flame ionization detector. The separation of the TAG critical pairs C48:0, C50:1, C52:2 and C54:3 [equivalent carbon number (ECN)=48] as well as C42:0, C48:3 and C54:6 (ECN=42) was accomplished without the aid of any interacting ion such as silver. Six TAG species were determined, but two of them, C54:3 (ECN=48) and C54:6 (ECN=42) were present to the extent of 74% or above of the total TAG content. The 4-desmethylsterols isolated from the unsaponifiable fractions were β-sitosterol (1135–1182 µg/g oil) as the main component followed by Δ5-avenasterol (925–1025 µg/g oil), and Δ7-avenasterol (615–809 µg/g oil). Stigmasterol, campesterol, and lanosterol were detected in small amounts.

Regiospecific analysis of neutral ether lipids by liquid chromatography/electrospray ionization/single quadrupole mass spectrometry: validation with synthetic compounds.

A reversed-phase high-performance liquid chromatography (HPLC) method with on-line electrospray ionization/collision-induced dissociation/mass spectrometry (ESI/CID/MS) is presented for the regiospecific analysis of synthetic reference compounds of neutral ether lipids. The reference compounds were characterized by chromatographic retention times, full mass spectra, and fragmentation patterns as an aid to clarify the regiospecificity of ether lipids from natural sources. The results clearly show that single quadrupole mass spectroscopic analysis may elucidate the regiospecific structure of neutral ether lipids. Ether lipid reference compounds were characterized by five to six major ions in the positive ion mode. The 1-O-alkyl-sn-glycerols were analyzed as the diacetoyl derivative, and showed the [M - acetoyl](+) ion as an important diagnostic ion. The diagnostic ions of directly analyzed 1-O-alkyl-2-acyl-sn-glycerols and 1-O-alkyl-3-acyl-sn-glycerols were the [M - alkyl](+), [M + H - H(2)O](+) and [M + H](+) ions. Regiospecific characterization of the fatty acid position was evident from the relative ion intensities, as the sn-2 species had relatively high [M + H](+) ion intensities compared with [M + H - H(2)O](+), whereas the reverse situation characterized the sn-3 species. Furthermore, corresponding sn-2 and sn-3 species were separated by the chromatographic system. However, loss of water was promoted as fatty acid unsaturation was raised, which may complicate interpretation of the mass spectra. The diagnostic ions of directly analyzed 1-O-alkyl-2,3-diacyl-sn-glycerols were the [M - alkyl](+), [M - sn-2-acyl](+) and [M - sn-3-acyl](+) ions. Regiospecific characterization of the fatty acid identity and position was evident from the relative ion intensities, as fragmentation of the sn-2 fatty acids was preferred to the sn-3 fatty acids; however, loss of fatty acids was also promoted by higher degrees of unsaturation. Therefore, both structural and positional effects of the fatty acids affect the spectra of the neutral ether lipids. Fragmentation patterns and optimal capillary exit voltages are suggested for each neutral ether lipid class. The present study demonstrates that reversed-phase HPLC and positive ion ESI/CID/MS provide direct and unambiguous information about the configuration and identity of molecular species in neutral 1-O-alkyl-sn-glycerol classes.

Variation of lipid classes among organs of the Northern krill Meganyctiphanes norvegica, with respect to reproduction

Lipid content and class in the Northern krill Meganyctiphanes norvegica (digestive gland, stomach, gonad, fat body, abdomen) was investigated and correlated with sex and reproductive stage. Ready to spawn females, have high lipid content in ovaries, while in males and spent females, the major site of lipid deposits was the digestive gland, followed by the fat body. These differences among spawning and spent females are indicative of strong interactions between the ovary and digestive gland and the ovary and fat body during vitellogenesis. Triacylglycerols (TAG) were the major neutral lipid class with high levels in the digestive gland. The major phospholipid was phosphatidylcholine (PC) particularly in the muscular tissue of the abdomen. Phosphatidyl-ethanolamine (PE) and –serine–inositol (PS–PI), were present at intermediate levels. Reproductive males were depleted in TAG and diacylglycerols (DAG) in the digestive gland, gonad and fat body, and had 4 times lower cholesterol in the gonad than ready to spawn females. Furthermore, ready to spawn females had in the ovary higher amounts of TAG, DAG and phospholipids (PC, PE, PS–PI) than spent females. Linear relationships between lipid content and main lipid class (TAG, PC, PE, PS–PI) in different fractions of males and ready to spawn females showed that: (1). TAG was stored for both sexes in all cephalothorax fractions with highest values in the digestive gland and ovary fluid; (2). PC was accumulated for both sexes in the fat body and the gonad with a higher slope for females, with the highest values in the ovary fluid and in the abdomen of males and that (3). PS–PI was stored only in the ovary and abdomen of mature females. These results are discussed in terms of the strategy developed by Meganyctiphanes norvegica to allocate lipids to the next generation for optimised embryogenesis.

Nondestructive Quantification of Neutral Lipids by Thin-Layer Chromatography and Laser-Fluorescent Scanning: Suitable Methods for “Lipidome” Analysis

A simple method for separation and quantification of neutral lipids was developed using thin-layer chromatography (TLC) and high-performance fluorescent scanning. Neutral lipid classes were separated using the double-developing TLC method and detected by rhodamine 6G and a laser-excited fluorescent scanner. The amount of lipids applied correlated with scanned intensity volume in a dose-dependent manner. The mass of each neutral lipid band was determined by comparing band intensities of unknown samples to dilution curves of authentic standards. After scanning the dye-sprayed TLC, acyl chain species of triglyceride (TG) extracted from TLC could be determined by gas chromatography. Using this method, we quantified the amounts of TG in mouse liver and found that the measured total mass of TG correlated with that obtained by enzymatic methods. Our method should provide the basic technique for “lipidome” analysis, designed to determine and compare total lipid classes and mass present in biological samples.

Lipids of Acorus calamus

Lipids of the leaves, stems, and rhizomes with roots ofAcorus calamusL. were investigated. The neutral-lipid classes and fatty-acid composition of neutral-, glyco-, and phospholipids were determined. The essential oil content in the total lipids of A. calamus was established by steam distillation.